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Communication Dans Un Congrès Année : 2020

Using direct long RNA sequencing to explore the whole mitochondrial transcriptome: model of the equine mitochondrial RNA

Résumé

Our interest is to analyse coding and non-coding mitochondrial transcriptomes in equine muscles to study their relationships with exercise capacity and myopathies. We are looking for undescribed alternative transcripts and long non-coding RNA. To find such molecules, we performed direct long RNA sequencing on RNA extracted from primary equine myoblasts. We used a direct RNA sequencing kit to build libraries that were sequenced using a minION sequencer connected to minIT device (Oxford Nanopore Technologies). We obtained 2,810 million reads (2.4 Gigabases). Quality controls revealed a N50 value of 1226 bases, a maximum length of RNA sequence of 30.9 kb, a median quality score of Q=8.8 (max 10), and 93% of the reads had a quality score over Q7. A total of 107,270 reads were mapped against the equine mitochondrial genome. The most expressed sense transcripts are ATP6>COX1>ND1>CYTB>tRNA-16S. The gene ND6 is expressed both in antisense and sense. The most expressed non-coding and antisense RNA are located on loci: D-loop>ND2>ND1>CYTB. The tRNAs are poorly expressed (10-1500 reads) and most of them are linked to the following genes. This direct long RNA sequencing method produced many long sequences with an acceptable quality score to investigate in details the native mitochondrial transcriptome.
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Dates et versions

hal-04510295 , version 1 (18-03-2024)

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  • HAL Id : hal-04510295 , version 1

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Sophie Dhorne-Pollet, Céline Robert, Eric Barrey. Using direct long RNA sequencing to explore the whole mitochondrial transcriptome: model of the equine mitochondrial RNA. 11th Mitochondria, World Mitochondria Society, Oct 2020, Berlin (DE), Germany. ⟨hal-04510295⟩
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